RESUMEN
Enzyme-linked immunosorbent assays have been widely used for diagnosis of FeLV and feline immunodeficiency virus (FIV) infections. Various ELISA kits for FeLV are available from several manufacturers. Although these tests are configured in a variety of formats, they are all direct antigen-detection systems for the viral core protein p27. On the other hand, ELISA for FIV exposure detects specific feline antibody to FIV. Basic immunoassay principles and the application of ELISA technology used in FeLV and FIV ELISA kits are described.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática , Virus de la Inmunodeficiencia Felina/aislamiento & purificación , Infecciones por Lentivirus/veterinaria , Virus de la Leucemia Felina/aislamiento & purificación , Leucemia Felina/diagnóstico , Animales , Anticuerpos Antivirales/sangre , Antígenos Virales/sangre , Gatos , Virus de la Inmunodeficiencia Felina/inmunología , Infecciones por Lentivirus/diagnóstico , Virus de la Leucemia Felina/inmunología , Valor Predictivo de las Pruebas , Juego de Reactivos para Diagnóstico/veterinariaAsunto(s)
Virus de la Inmunodeficiencia Felina/inmunología , Infecciones por Lentivirus/veterinaria , Virus de la Leucemia Felina/inmunología , Leucemia Felina/epidemiología , Factores de Edad , Animales , Anticuerpos Antivirales/sangre , Antígenos Virales/sangre , Castración/veterinaria , Gatos , Ensayo de Inmunoadsorción Enzimática , Femenino , Infecciones por Lentivirus/complicaciones , Infecciones por Lentivirus/epidemiología , Leucemia Felina/complicaciones , Masculino , Prevalencia , Factores de Riesgo , Factores Sexuales , Estados Unidos/epidemiologíaRESUMEN
The feline immunodeficiency virus (FIV) is a recently identified feline lentivirus that has been found at significant levels in domestic cat populations worldwide. A microdilution plate format, monoclonal antibody-based enzyme-linked immunosorbent assay was developed for the detection of the FIV group-associated antigen (gag) designated p24. Assays of serially diluted samples containing disrupted virus showed that the assay had a sensitivity limit of approximately 0.2 ng/ml for FIV p24. The assay was approximately eightfold more sensitive than the assay for viral reverse transcriptase activity when it was tested with diluted tissue culture samples. A qualitative confirmation assay by standard antibody inhibition techniques was coupled to the screening test methodology. The test was used to detect and confirm the presence of virus in cultured feline lymphocytes from infected animals.
Asunto(s)
Antígenos Virales/análisis , Ensayo de Inmunoadsorción Enzimática , Retroviridae/inmunología , Proteínas del Núcleo Viral/inmunología , Animales , Anticuerpos Monoclonales , Anticuerpos Antivirales , Gatos , Estudios de Evaluación como Asunto , Infecciones por Retroviridae/diagnósticoRESUMEN
The feline T-cell lymphotropic lentivirus (feline immunodeficiency virus) is a recently described feline-specific retrovirus that can produce chronic immunodeficiency-like disorders in cats. A microdilution plate format enzyme-linked immunosorbent assay has been developed to detect the presence of antibody to the virus in feline serum or plasma. Temporal studies performed with experimentally infected animals show that seroconversion can be demonstrated 3 to 4 weeks after exposure to the virus. Results of a serosurvey (n = 1,556 samples) indicate that infection is fairly common in both clinic (5.2%) and sick cat (15.2%) populations. Western blot (immunoblot) and sodium dodecyl sulfate radioimmunoprecipitation assays were developed to confirm microdilution plate test results and to identify peptides specific for the feline immunodeficiency virus. All microdilution plate test positive results and selected negative results were confirmed by one or both of these procedures. These data demonstrate that this microassay plate enzyme-linked immunosorbent assay is a very sensitive and specific test for detection of antibody to the feline immunodeficiency virus.
Asunto(s)
Anticuerpos Antivirales/análisis , Enfermedades de los Gatos/diagnóstico , Ensayo de Inmunoadsorción Enzimática , Infecciones por Retroviridae/veterinaria , Retroviridae/inmunología , Animales , Western Blotting , Gatos , Electroforesis en Gel de Poliacrilamida , Pruebas de Precipitina , Valor Predictivo de las Pruebas , Infecciones por Retroviridae/diagnósticoRESUMEN
Treatment of mouse mammary glands with a high concentration of 7,12-dimethylbenzo(a)anthracene in whole organ culture was reported by Banerjee et al. to transform foci of lobuloalveoli to a hormone-independent state, and to give rise to mammary hyperplastic outgrowths and adenocarcinomas in vivo. In the present study using the identical system, mammary glands of BALB/c mice were exposed to 7,12-dimethylbenzo(a)anthracene or N-2-fluorenylacetamide at low concentrations that bring about maximal incidences of the hormone-independent hyperplastic lobuloalveolar lesions with minimal cytotoxicity. After morphological development of the lobuloalveoli in culture, the glands were enzymatically dissociated into cells and inoculated into gland-free inguinal mammary fat pads of syngeneic mice bearing pituitary gland implants during the initial 8 weeks. After 11 months, fragments of the resultant mammary outgrowths from each mouse were implanted into the gland-free inguinal mammary fat pads of 3 syngeneic mice (not bearing pituitary gland supplements) and were permitted to grow for another 11 months. Mammary outgrowths from the primary and secondary implants were neither neoplastic, anaplastic, nor dysplastic. Also, no hyperplasia in any mammary outgrowth could be attributed to the action of either carcinogen, especially when outgrowths were compared with contralateral outgrowths that arose from the control glands exposed to dimethyl sulfoxide (solvent of the carcinogens) in culture and/or with untreated thoracic mammary glands of the same hosts. One interpretation of these findings is that the hormone-independent, hyperplastic alveolar lesions may not be an appropriate in vitro marker of oncogenic transformation by chemical carcinogens in culture. The great variety of procarcinogens and activated carcinogens that bring about this lesion in vitro and its morphological similarity to presumptive mammary preneoplastic lesions in vivo weigh against this interpretation. A second hypothesis is that high concentrations of procarcinogens, despite their considerable cytotoxicity, complete a multistep process of oncogenic transformation in surviving mammary epithelium, whereas low concentrations optimized to produce the lesions in maximal number do not.
Asunto(s)
2-Acetilaminofluoreno/toxicidad , 9,10-Dimetil-1,2-benzantraceno/toxicidad , Benzo(a)Antracenos/toxicidad , Glándulas Mamarias Animales/efectos de los fármacos , Neoplasias Mamarias Experimentales/inducido químicamente , Animales , Relación Dosis-Respuesta a Droga , Hiperplasia/inducido químicamente , Glándulas Mamarias Animales/patología , Glándulas Mamarias Animales/trasplante , Ratones , Ratones Endogámicos BALB C , Técnicas de Cultivo de ÓrganosAsunto(s)
Carcinógenos/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Glándulas Mamarias Animales/efectos de los fármacos , Neoplasias Mamarias Experimentales/inducido químicamente , Virus del Tumor Mamario del Ratón/efectos de los fármacos , Activación Viral/efectos de los fármacos , 2-Acetilaminofluoreno/farmacología , 9,10-Dimetil-1,2-benzantraceno/farmacología , Animales , Transformación Celular Viral , Células Cultivadas , ADN Viral/biosíntesis , Femenino , Neoplasias Mamarias Experimentales/microbiología , Virus del Tumor Mamario del Ratón/genética , Ratones , Ratones Endogámicos , ARN Viral/análisis , Proteínas Virales/análisisAsunto(s)
Bucladesina/farmacología , Diferenciación Celular/efectos de los fármacos , Factor de Crecimiento Epidérmico/farmacología , Glándulas Mamarias Animales/fisiología , Prostaglandinas/farmacología , Animales , Caseínas/biosíntesis , Células Cultivadas , AMP Cíclico/metabolismo , Femenino , Glándulas Mamarias Animales/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Peso MolecularRESUMEN
The mouse mammary gland in serum-free whole organ culture can be manipulated hormonally to undergo one complete physiological cycle consisting of lobuloalveolar development, functional differentiation and regression, mimicking processes that occur in vivo. A second cycle has not previously been achieved in vitro. The present study has identified a specific requirement for epidermal growth factor (EGF) in the morphological development of mammary lobuloalveoli, allowing two complete cycles of development and regression in culture.